臺灣博碩士論文加值系統

糖尿病是一種代謝性疾病，其病徵是在於患者的血糖長期高於標準值。目前全世界至少有1.5億人罹患糖尿病，顯示出糖尿病已嚴重威脅國民的健康，而目前糖尿病患者中約90 %的患者屬於第二型的糖尿病。近年來已有研究顯示指出，二型糖尿病會因肥胖所產生的發炎因子，如:腫瘤壞死因子-α(TNF-α)、介白素-6(IL-6)、介白素-1β(IL-1β)…等，導致發炎反應，進而造成胰島素阻抗。在正常的胰島素訊號通路下，胰島素會與胰島素接受體接合，經磷酸化胰島素受體受質(IRS1/2/3)後，再往下游的脂醯醇3激酶(PI3K)及蛋白激酶B(Protein kinase B；PKB，又稱Akt)傳遞訊號，最終使葡萄糖轉運蛋白4(GLUT4)位移到細胞膜上，運送葡萄糖進入細胞內利用。而蛋白酪氨酸磷酸酶1B(PTP1B)是胰島素信號傳導途徑的負調節物，在胰島素阻抗的情況下，PTP1B會加劇高血糖的現象。常用來作為中醫臨床上的降血糖藥的青錢柳(Cyclocarya paliurus)，其富含多種化學成分，除了可以降三高，亦被證實具有抗氧化壓力、抗發炎、保護肝臟及腎臟的功能。本研究主要探討青錢柳葉萃取物(C. paliurus Extract, CPE)針對調節血糖與發炎之功效進行機轉研究。首先由RIN-m5F細胞增生率試驗中發現:以CPE 50.0 μg/mL作用24小時後，可達到最佳的增生效果(71.0 ％)。在小鼠調節血糖試驗中，經連續管餵各劑量之CPE 28天後，CPE中劑量組(200.0 mg/mL)可顯著提升總蛋白質(14.89 %)、降低空腹血糖值(65.48 %)、三酸甘油脂(28.30 %)及總膽固醇(31.32 %)；增加IRS2(108.43 %)、Akt1/2/3(93.18 %)及GLUT4(140.22 %)的表現量，並與STZ組達到顯著性統計差異(p< 0.05)。另由小鼠抗發炎實驗中發現: CPE高劑量(25.0 mg/mL)組經12、24小時後，相較於TPA組，分別抑制了26.92 %及27.18 %，其抑制效果更優於陽性對照藥(Indomethacin)。在PTP1B抑制試驗中，隨著CPE劑量增加，PTP1B的含量也隨之減少，在CPE高劑量(5.0 mg/mL)下有最低濃度(0.001 ± 0.08 nmol)，顯示CPE濃度越高抑制的效果越好。綜合上述實驗結果，CPE可藉由調節生化指標以及加強胰島素信號的傳遞，改善糖尿病鼠高血糖的現象，並藉由抗發炎反應減輕糖尿病鼠的胰島素阻抗，此外也可透過抑制PTP1B的作用，增強胰島素訊號的傳導。由此得知:CPE確實可藉由多方面的作用來調節血糖，而有益於糖尿病的恢復。

Diabetes is a metabolic disease. Diabetes is characterized by the blood sugar of the patient is over the standard value for a long time. At least 150 million people worldwide suffer from diabetes. This incident shows that diabetes has seriously threatened the health of the people. Currently, about 90% of patients with diabetes are classified as type 2 diabetes. In recent years, studies have shown that type 2 diabetes causes inflammatory factors such as TNF-α, IL-6 and IL-1β...etc, which produce an inflammatory response that in turn causes insulin resistance. In the normal insulin signaling pathway, insulin binds to the insulin receptor and phosphorylates IRS1/2/3, PI3K and Akt. Eventually, GLUT4 is displaced onto the cell membrane and glucose is transported into the cell for utilization. PTP1B is a negative regulator of the insulin signaling pathway. PTP1B exacerbates hyperglycemia in the case of insulin resistance. Cyclocarya paliurus is commonly used as a clinical hypoglycemic agent in traditional Chinese medicine. It is rich in a variety of chemical ingredients, which not only can lower the three highs, oxidative stress and inflammatory response. This study focused on the effects of Cyclocarya paliurus leaf-extract (CPE) on the regulation of blood sugar and inflammation. First of all discovered by the test of RINm5F cell proliferation, after addition CPE 50.0 μg/mLand 24 hours, the optimal proliferation effect (71.0%) was achieved. In the mouse of blood sugar regulation test, after continuous administration of CPE for 28 days, The CPE medium dose group (200.0 mg/mL) significantly increased Total-protein (14.99%), decreased Glucose AC (65.48%), Triglyceride (28.33%) and Total-cholesterol (31.32%); increased the performance of IRS2 (108.43%), Akt1/2/3 (93.18%) and GLUT4 (140.22%), and significant statistical differences (p<0.05) compared with the STZ group. In addition, compared with the TPA group, the reduction of ear edema was 26.9 % and 27.2 % respectively, after administration of high dose CPE (25.0 mg/mL) for 12 and 24 hours in the anti-inflammatory experiment of mice. In addition, compared with the TPA group, the reduction of ear edema was 26.9 % and 27.2 % respectively, after administration of high dose CPE ( 25.0 mg/mL ) for 12 and 24 hours in the anti-inflammatory experiment of mice, and the inhibitory effect is better than the positive control Medicine (Indomethacin). In the PTP1B inhibition test, the PTP1B content decreased with the higher dose. At the high dose of CPE (5.0 mg/mL), it has the lowest concentration (0.001 ± 0.08 nmol). It shows that the higher CPE concentration has the better inhibitory effect. Based on the above experimental results, CPE can improve the hyperglycemia of diabetic patients by regulating biochemical indicators and enhancing the transmission of insulin signals, and reduce the insulin resistance of patients by anti-inflammatory reaction. In addition, it can enhance insulin signal transmission by inhibiting the action of PTP1B. CPE can regulate blood sugar in many ways, which is beneficial to the recovery of diabetes.

摘 要 ................................................... I
Abstract .............................................. III
誌謝 .................................................... V
目 錄 .................................................. VI
圖 次 目 錄 ............................................. IX
縮 寫 表 ................................................ X
第壹章 緒言 ............................................. 1
實驗架構 ................................................ 2
第貳章 文獻回顧 .......................................... 3
(一) 青錢柳簡介及藥理活性 ................................. 3
(二) 糖尿病簡介 .......................................... 7
(三) 發炎簡介 ........................................... 13
第參章 材料與方法......................................... 18
(一) 實驗相關儀器及藥品 .................................. 18
(二) 材料製備 ........................................... 22
(三) RIN-m5F細胞增生率試驗方法 ........................... 25
(四) CPE調節小鼠血糖試驗方法 ............................. 27
(五) CPE對TPA誘導小鼠耳朵腫脹試驗方法 ..................... 30
(六) PTP1B抑制試驗方法 ................................... 31
(七) 統計分析 ........................................... 31
第肆章 結果 ............................................. 32
(一) CPE對大鼠胰臟RIN-m5F細胞增生率試驗之結果 .............. 32
(二) CPE調節小鼠血糖試驗之每週體重、攝食量及攝水量結果 ....... 38
(三) CPE調節小鼠血糖試驗之生化檢測結果 ..................... 43
(四) CPE調節小鼠血糖試驗之機轉分析結果 ..................... 51
(五) CPE對TPA誘導小鼠耳朵腫脹試驗方法結果 .................. 57
(六) CPE對PTP1B之抑制實驗結果 ............................. 60
第伍章 討論 .............................................. 62
第陸章 結論 .............................................. 66
參考文獻 ................................................ 67

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